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. 2015 Nov 22;6(39):41550–41565. doi: 10.18632/oncotarget.6355

Figure 4. ASPP1/2 co-depletion causes SAC hyperactivation.

Figure 4

a. Localization of Mad1, Mad2 and Mps1 in ASPP1/2 co-depleted HeLa cells. HeLa cells were transfected with control or ASPP1/2 siRNAs for 48 hr, and treated with nocodazole for 12 hr and then released into fresh media for 1-2 hr before fixation. Cells were stained with antibodies against the indicated SAC proteins (red), together with kinetochores (CREST, green) and DNA (blue). The figures show confocal images of cells at prometaphase and metaphase. Insets are magnified images of the boxed areas. Scale bar = 10 μm. b. Quantification of the fluorescence intensity of the SAC proteins normalized to the fluorescence intensity of CREST staining are shown. For quantifications, ∼30 mitotic cells were measured for each experiment and condition. Error bars, SEM *p<0.01 from triplicates. c. WB analyses of cell lysates prepared from control and ASPP1/2 co-depleted HeLa cells using the indicated antibodies.